This project aims to identify small molecules to investigate the Rcs pathway. Supported by an NIH Intramural DDIR Innovation Award, the team is pursuing screening of more than 50,000 natural products product extracts from the Natural Products Discovery Institute (NPDI) as a reservoir for identifying new antibiotics. This work should provide a much better understanding of the regulation of capsule synthesis in Klebsiella, possible lead compounds for development of novel antibiotics aimed to change bacterial behavior rather than killing the bacteria, and provide structural insight into some of the unique aspects of the capsule regulatory cascade, relevant to a broad group of gram-negative bacteria. During this period, the collaborative team has successfully optimized a 1536-well high-throughput assay for small molecule screening, which was utilized to screen a number of NCGC's small molecule collections, including a natural product extract collection. Several compounds capable of activating the Rcs pathway have been identified, and these hits have been cherrypicked for further investigation using in vivo models.